Andersson J, Nagy S, Groth C G, Andersson U
Department of Immunology, Arrhenius Laboratories for Natural Sciences, Stockholm University, Sweden.
Immunology. 1992 Jan;75(1):136-42.
Mononuclear cells obtained from human blood were mitogen or antigen activated in vitro in the presence or absence of FK506 or cyclosporin A (CsA). Cytokine production was studied at a single-cell level by ultraviolet (UV) microscopy of fixed permeabilized cells using cytokine-specific monoclonal antibodies (mAb). Phenotypic characterization of the monokine-producing cells was achieved by two-colour immunofluorescent staining. Cytokine production after antigen activation with Staphylococcus aureus enterotoxin A (SEA) was significantly reduced. FK506 or CsA inhibited SEA-induced tumour necrosis factor-alpha (TNF-alpha) production both in monocytes (P less than 0.01) and in lymphocytes (P less than 0.001), at a drug concentration of 1-25 ng/ml for FK506 and 100-500 ng/ml for CsA. Lymphocyte synthesis of interleukin-2 (IL-2), interferon-gamma (IFN-gamma) and TNF-beta after SEA activation was also significantly reduced by either of the drugs. In contrast, endotoxin-induced monokine production (TNF-alpha and IL-6) after lipopolysaccharide (LPS) stimulation was unaffected by FK506 or CsA even when added in concentrations as high as 1000 ng/ml. When the cells were stimulated by phorbol ester (phorbol 12-myristate 13-acetate, PMA) plus calcium ionophore (ionomycin), FK506 and CsA inhibited, in a dose-dependent manner, the production of IL-2, IL-4, IL-5, IFN-gamma and TNF-alpha. The 50% inhibitory concentration (IC50) for FK506 or CsA on the cellular synthesis of the various cytokines varied between 0.6 and 1.0 ng/ml and 20 and 60 ng/ml, respectively. Further stimulation by addition of anti-CD28 mAb to the cultures resulted in an augmented IL-2 and IFN-gamma production which was resistant to both FK506 and CsA. This report delineates extensive similarities between the two drugs in mechanisms of immunosuppression by blockade of identical interleukin production. Depending on the mode of cell activation the two drugs inhibited not only cytokine production in lymphocytes but also antigen-induced monokine (TNF-alpha) production in macrophages, although the optimal immunomodulatory effect of FK506 was achieved at a concentration approximately 50-fold lower than that of CsA.
从人血液中获取的单核细胞在体外有丝分裂原或抗原激活时，存在或不存在FK506或环孢菌素A（CsA）。通过使用细胞因子特异性单克隆抗体（mAb）对固定通透细胞进行紫外线（UV）显微镜检查，在单细胞水平上研究细胞因子的产生。通过双色免疫荧光染色对产生单核因子的细胞进行表型鉴定。用金黄色葡萄球菌肠毒素A（SEA）进行抗原激活后，细胞因子的产生显著减少。FK506或CsA在药物浓度为1 - 25 ng/ml（FK506）和100 - 500 ng/ml（CsA）时，抑制SEA诱导的单核细胞（P<0.01）和淋巴细胞（P<0.001）中肿瘤坏死因子-α（TNF-α）的产生。SEA激活后，两种药物也显著降低淋巴细胞中白细胞介素-2（IL-2）、干扰素-γ（IFN-γ）和肿瘤坏死因子-β的合成。相比之下，脂多糖（LPS）刺激后内毒素诱导的单核因子产生（TNF-α和IL-6）不受FK506或CsA的影响，即使添加浓度高达1000 ng/ml。当细胞用佛波酯（佛波醇12 - 肉豆蔻酸酯13 - 乙酸酯，PMA）加钙离子载体（离子霉素）刺激时，FK506和CsA以剂量依赖性方式抑制IL-2、IL-4、IL-5、IFN-γ和TNF-α的产生。FK506或CsA对各种细胞因子细胞合成的50%抑制浓度（IC50）分别在0.6至1.0 ng/ml和20至60 ng/ml之间变化。向培养物中添加抗CD28 mAb进一步刺激导致IL-2和IFN-γ产生增加，且对FK506和CsA均有抗性。本报告描述了两种药物在通过阻断相同白细胞介素产生进行免疫抑制机制方面的广泛相似性。根据细胞激活方式，两种药物不仅抑制淋巴细胞中的细胞因子产生，还抑制巨噬细胞中抗原诱导的单核因子（TNF-α）产生，尽管FK506的最佳免疫调节作用在比CsA低约50倍的浓度下实现。
